title: Dynamics of ESCRT protein recruitment during retroviral assembly.
abstract: The ESCRT (endosomal sorting complex required for transport) complexes and associated proteins mediate membrane scission reactions, such as multivesicular body formation, the terminal stages of cytokinesis and retroviral particle release. These proteins are believed to be sequentially recruited to the site of membrane scission, and then complexes are disassembled by the ATPase Vps4A. However, these events have never been observed in living cells, and their dynamics are unknown. By quantifying the recruitment of several ESCRT and associated proteins during the assembly of two retroviruses, we show that Alix progressively accumulated at viral assembly sites, coincident with the accumulation of the main viral structural protein, Gag, and was not recycled after assembly. In contrast, ESCRT-III and Vps4A were transiently recruited only when the accumulation of Gag was complete. These data indicate that the rapid and transient recruitment of proteins that act late in the ESCRT pathway and carry out membrane fission is triggered by prior and progressive accumulation of proteins that bridge viral proteins and the late-acting ESCRT proteins.
authors.affiliation: Aaron Diamond AIDS Research Center, Laboratory of Retrovirology, The Rockefeller University, New York, 10065 New York, USA.
authors.forename: Nolwenn
authors.initials: N
authors.lastname: Jouvenet
authors.forename: Maria
authors.initials: M
authors.lastname: Zhadina
authors.forename: Paul D
authors.initials: PD
authors.lastname: Bieniasz
authors.forename: Sanford M
authors.initials: SM
authors.lastname: Simon
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics / Q000378:metabolism*; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D004860:Infectious Anemia Virus, Equine / Q000502:physiology / Q000648:ultrastructure; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000502:physiology* / Q000648:ultrastructure; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D015683:Gene Products, gag; C117020:PDCD6IP protein, human; D011993:Recombinant Fusion Proteins; D025262:Vacuolar Proton-Translocating ATPases; D000074183:ATPases Associated with Diverse Cellular Activities; C490305:VPS4A protein, human